Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.

The N-terminal and C-terminal regions of shot interact with each other.

The basis of this technique is that when tagged proteins interact, the complementary fragments of Venus are brought into close proximity and reconstitute a full-length, fluorescent protein.

shot forms intramolecular interactions in cis, but not intermolecular interactions in trans. Reconstitution of Venus fluorescence was observed only when the complementary Venus fragments were present in cis at opposite ends of the same shot molecule. When complementary halves of shot were co-expressed, no fluoresence complementation could be detected.

The flexible Rod domain of shot is required for intramolecular interaction.

Source was live S2 cells; proteins produced from transfected construct.